human lung tissue sections  (R&D Systems)

Journal: Respiratory Research

Article Title: Monoclonal enolase-1 blocking antibody ameliorates pulmonary inflammation and fibrosis

doi: 10.1186/s12931-023-02583-3

Figure Lengend Snippet: ENO1 is upregulated in fibrotic lungs from human and bleomycin-treated mice. (A, B) IHC staining of ENO1 was performed in commercially available human normal (n = 3) and fibrosis (n = 3) lung FFPE sections. (C, D) After intratracheal injection of bleomycin (BLM, 3 mg/kg) (n = 7) or PBS vehicle control (Sham) (n = 4), the mouse lungs were harvested on day 21 for preparation of FFPE sections and subjected to IHC staining of ENO1. Representative pictures (A, C) and quantitative results of ENO1-stained positive areas were shown (B, D) . After intratracheal injection of bleomycin (BLM, 3 mg/kg) (n = 4) or PBS vehicle control (Sham) (n = 4), the mouse lungs were harvested on day 21 for preparation of lysates and subjected to Western blotting for ENO1 (E) and the relative densitometry was shown below the representative blot after GAPDH normalization. Cropped blots were shown, and supplementary Fig. presented the full-length blots. Quantitative results were shown by fold change after bleomycin treatment (F) . Scale bar, 100 µM. * P < 0.05, ** P < 0.01. (A, B) One experiment was performed and each picture or data point was from one human subject. (C-F) Data were representative for two independent experiments. Each picture, data point, or protein band was from one mouse except ( F ) was shown as mean ± SD

Article Snippet: Three human fibrotic lung FFPE sections (#CS701530, #CS702702, #CS703355) were obtained from OriGene (Rockville, MD, USA).

Techniques: Immunohistochemistry, Injection, Control, Staining, Western Blot

Journal: Respiratory Research

Article Title: Monoclonal enolase-1 blocking antibody ameliorates pulmonary inflammation and fibrosis

doi: 10.1186/s12931-023-02583-3

Figure Lengend Snippet: Blocking ENO1 ameliorates lung fibrosis in bleomycin-treated mice. After intratracheal injection of 3 mg/kg bleomycin (BLM) (day 0), mice were treated with ENO1 Ab HL217 (10 mg/kg) intravenously on a 6-day interval from day 1. Mouse lungs were harvested on day 14 or 21 for preparation of FFPE sections and subjected to H&E staining (A) and Masson’s trichrome staining (B) . Representative pictures on day 21 (A, B) and quantitative results of Ashcroft score on day 21 (C) and inflammation score on day 14 (D) were shown. Body weight change (E) , ratio of lung weight versus body weight (F) , collagen content of the lungs (G) , and levels of TGF-β in BALF (H) were shown. Scale bar, 100 µM. * P < 0.05, ** P < 0.01, *** P < 0.001. Data were representative for two independent experiments. Each picture or data point was from one mouse except (E) was shown as mean ± SEM

Article Snippet: Three human fibrotic lung FFPE sections (#CS701530, #CS702702, #CS703355) were obtained from OriGene (Rockville, MD, USA).

Techniques: Blocking Assay, Injection, Staining

Journal: Cell Reports Methods

Article Title: Chromatin accessibility profiling of targeted cell populations with laser capture microdissection coupled to ATAC-seq

doi: 10.1016/j.crmeth.2023.100598

Figure Lengend Snippet: Assessing the chromatin accessibility of scattered tissue cell populations (A) Schematic representation of the LCM-ATAC-seq workflow in human left lobe (LL) lung parenchyma samples. L1: lower LL; L2: upper LL. (B) Representative immunofluorescence PU.1 staining of an L2 section from donor 4 (40× magnification). (C) Overall TSS enrichment profile of analyzed human LCM-ATAC-seq samples. (D) FSD across analyzed human samples. (E) SPI1 locus accessibility in normalized PU.1 aggregate track versus lung bulk ENCODE reference (human lung bulk: ENCSR647AOY dataset, ENCFF210HIS.bigwig). (F) SFTPB locus accessibility in normalized PU.1 aggregate bigwig track versus lung bulk ENCODE reference (human lung bulk: ENCSR647AOY dataset, ENCFF210HIS.bigwig). (G) One-tailed fast gene set enrichment analysis for signatures PU1_Q6 (PU.1 transcription factor targets) and GO_BP_MYELOID_CELL_DIFFERENTIATION on PU.1 targeted cells rank of chromatin accessibility. FF, fresh frozen.

Article Snippet: Human lung tissue sections were first incubated with 100 μL blocking buffer consisting of 2% normal goat serum (08642921, MP biomedicals) diluted 1:5 in PBST for 45 min at room temperature.

Techniques: Immunofluorescence, Staining, One-tailed Test, Cell Differentiation

Journal: Cell Reports Methods

Article Title: Chromatin accessibility profiling of targeted cell populations with laser capture microdissection coupled to ATAC-seq

doi: 10.1016/j.crmeth.2023.100598

Figure Lengend Snippet:

Article Snippet: Human lung tissue sections were first incubated with 100 μL blocking buffer consisting of 2% normal goat serum (08642921, MP biomedicals) diluted 1:5 in PBST for 45 min at room temperature.

Techniques: Recombinant, dsDNA Assay, Software